Enhancing acid stability of citral through internal structure modulation in nanostructured lipid carriers with solid lipids and phospholipids.

In this current study, the internal structure of nanostructured lipid carriers was modulated by phospholipids (lecithin PC, hydrogenated soybean phospholipid HPC) and solid lipids to achieve stable encapsulation of citral. The presence of high melting point HPC could construct α-crystalline type with more lattice defects and effectively inhibit ß-ization. The HPC group could maintain the particle size at 155.9-186.9 nm, the polydispersity index (PDI) at 0.182-0.321, the Zeta potential at -57.58 mV to -49.35 mV and the retention rate of citral at 91.33-98.49 % in the acidic environments of 2 mM and 20 mM hydrochloric acid solutions. The recrystallization index (RI) of NLC increased with the number of solid lipid ester bonds (from 3.57 % to 16.58 % in the PC group and from 0.82 % to 12.47 % in the HPC group). The results illustrated that the number of solid lipid ester bonds and the melting point of phospholipids affected crystallinity of the lipid matrix and thus the stability of encapsulated citral. Hydrogenated phospholipid with high melting points was more beneficial in stabilizing citral. The present study improved the acidic stability of citral and provided a new thought for the application of citral in acidic beverages.

Infection of grass carp reovirus induced the expressional suppression of pro-viral Fibulin-4 in host cells.

Fibulin-4 is not only involved in connective tissue development and elastic fiber formation, but also plays critical neoplastic roles in tumor growth by activating Wnt/ß-Catenin signaling in human. Recently, Fibulin-4 was shown to associate with grass carp reovirus (GCRV) outer capsid proteins and might relate to viral hemorrhagic disease in grass carp Ctenopharyngodon idella. Here, we monitored the expression pattern of Fibulin-4 during the infection course of GCRV at both translational and transcriptional levels, and found that Fibulin-4 was significantly suppressed upon the viral challenge in grass cap GCO cells. Over expression of Fibulin-4 was achieved by transduction of pEGFP-Fibulin-4 plasmids into GCO cells, which was confirmed by both Western blot and Real time RT-PCR analysis. In GCO cells with over-expression of Fibulin-4, significantly increase of viral protein synthesis and progeny virus production was detected. Our study indicated that Fibulin-4 displayed pro-viral function and was inhibited during viral challenge. Thus, repression of Fibulin-4 expression seemed to be involved in anti-viral response in grass carp Ctenopharyngodon idella.

Repression of SUMOylation pathway by grass carp reovirus contributes to the upregulation of PKR in an IFN-independent manner.

SUMOylation, a post-translational modification, is involved in interaction between hosts and viruses, and participates in diverse cellular processes including inflammatory responses and innate immunity. Here, we investigated the interaction between reovirus infection and the cellular SUMOylation machinery using grass carp reovirus (GCRV) as a model. Full-length cDNAs of grass carp SUMO-1 and SUMO-2 were obtained and phylogenetic analysis indicated that they shared high homology with those of higher vertebrates. The two modifiers and SUMO conjugating enzyme 9 (Ubc9) were ubiquitously expressed in all tested tissues of grass carp. During GCRV infection in CIK cells, transcriptional expressions of SUMO1/2 and Ubc9 were significantly inhibited; while UV-inactivated GCRV failed to inhibit the expression of the three molecules, which suggested that SUMOylation system was suppressed during viral replication. In CIK cells treated with inhibitor 2-D08 for SUMOylation, GCRV replication was not interfered; however, transcriptional analysis of immune genes involved in anti-viral interferon (IFN) response indicated that IRF2 and PKR were significantly up-regulated in CIK cells treated with inhibitor in contrast to IRF1, IRF7 and IFNI. Furthermore, 2-D08 treatment coupled with GCRV challenge resulted in higher IRF2 and PKR level during infection in comparison to those of CIK cells infected with GCRV only. These results indicated that inhibition of SUMOylation should result in the induction of PKR via IFN-independent manner, and both IFN-signaling and IFN-independent signaling seemed to involve in the upregulation of PKR during the process of GCRV infection. Repression of SUMOylation by GCRV might represent a cellular antiviral mechanism.